It is estimated that 100 million men and women are affected by urinary incontinence (Ul). The prevalence of Ul is generally higher in women than in men, women being between two (older age groups) and four times younger and middle-aged) more likely to be incontinent than men. In the past 4 years, supported by a SCOR grant (P50 DK64538, PI: Brown, J), we have developed a new 70-mer oligonucleotide glass microarray to examine and identify genes associated with stress urinary incontinence (SUI) in our rat model. We also identified several growth factors that promote nerve growth from the female pelvic ganglion-the origin of cholinergic neurons innervating bladder and urethra. We changed our original proposal from using organ-specific acellular matrix to growing autologus adipose tissue derived stem cells (ADSCs) onto the acellular matrix for the treatment of SUI and obtained very encouraging results. We therefore propose to study the in vivo tissue engineering and molecular mechanisms involved so that we can develop a method to nstruct and regulate the differentiation of ADSCs shortly before and after implantation. To better integrate with the research in project 1 (Dr. Thorn's group) and project 3 (Dr. Brown's group), we also developed a comprehensive urodynamic test to study the voiding function of a type 2 diabetic model-the Zucker Diabetic Fat (ZDF) rat. We have since discovered that these rats developed a whole spectrum of voiding dysfunction from overactive bladder to urinary retention similar to the symptoms of women with "diabetic cystopathy".We propose to continue our research to examine the natural course and reversibility of diabetic voiding dysfunction in the diabetic ZDF rat model and identify the molecular mechanisms involved. Our long-term goal is to better understand stem cell biology and diabetic voiding dysfunction so that the information can be "translated" to better prevention, diagnosis and treatment of stress urinary incontinence and diabetic voiding dysfunction in women. To achieve this overall goal we have set three specific aims for our proposed study. Specific Aim 1: To study the in vivo tissue engineering and molecular mechanism of adipose derived stem cell (ADSC) differentiation Specific Aim 2: To apply ADSCs for the prevention and correction of stress urinary incontinence in our rat model Specific Aim 3: To study the molecular mechanism of diabetic voiding dysfunction in a type 2 diabetic rat model